Method for the rapid diagnosis of targets in human body fluids using undiluted samples

ABSTRACT

More particularly, the present invention relates to a method for the detection of a target, e.g. pathogen in a human body fluid wherein a body fluid sample is collected with a swab member.

REFERENCE TO RELATED APPLICATIONS

This is a divisional patent application of copending application Ser.No. 11/698,053, filed Jan. 26, 2007, entitled “Method for the RapidDiagnosis of Targets in Human Body Fluids” and copending applicationSer. No. 11/052,748, filed Feb. 9, 2005, entitled “Method for the RapidDiagnosis of Targets in Human Body Fluids”, which claimed one or moreinventions which were disclosed in U.S. Provisional Application No.60/542,303, filed Feb. 9, 2004, entitled “Method for the Rapid Diagnosisof Targets in Human Body Fluids”. The benefit under 35 USC §119(e) ofthe United States provisional application is hereby claimed, and theaforementioned applications are hereby incorporated herein by reference.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to a method for the detection of targets,e.g. pathogens and/or allergy-associated components in a human bodyfluid wherein a body fluid sample is collected with a swab member. Thesamples are transferred from the swab member to a sample analysisdevice, on which an analysis of the targets, e.g. by immunochemical orenzymatic means can take place. The test result may be displayed withina short period of time and can be directly read out by the user.Further, a test kit for carrying out the method of the invention isprovided.

BACKGROUND OF THE INVENTION

Rapid, point-of-care analysis is becoming increasingly important in thediagnosis and treatment of various viral and other pathogenicmicrobiological agents (bacteria, others). Especially in the acutestatus of a infectious disease medical doctors have a need for immediatedetection of the causal agent for the symptoms observed.

Prior art discloses a rapid assay for HIV specific antibodies in salivasamples. A saliva sample is gained by means of a sampling stick. Thesaliva sample is diluted in a sample buffer and a lateral flowimmunoassay is dipped into the diluted saliva sample [U.S. Pat. No.5,714,341].

German Patent Nr. DE19622503 suggests to apply lateral flow immunoassaysfor the detection of illegal narcotics in saliva or sweat.

Conjunctivitis, commonly known as red eye or pink eye, may be caused byseveral different agents including viruses, bacteria and allergens.Different etiologies require different treatments. Infectiousconjunctivitis is typically contagious. Conjunctivitis is generallydiagnosed clinically, by gross examination, and (during a routine eyeexam) slit lamp biomicroscopy. This method does not provide informationon the specific infectious agent. If specific (pathogen typing)diagnosis is necessary, swabs of the inferior formix are sent forlaboratory analysis to determine the type of pathogen. The preferredmethods for laboratory analysis are cell culture with confirmatorydirect immunofluorescence, ELISA or PCR. The disadvantage of thisdiagnostic strategy is that laboratory analysis needs typically betweentwo and ten days, utilizes complex diagnostic equipment, and may requiretechnical skill in both performing and interpreting results. This timeperiod is problematic for a proper treatment of potentially infectiousforms of conjunctivitis that cannot be specifically classified/connectedwith a certain pathogenic agent.

A publication by Uchio et al. (Opthalmology 104 (1997), 1294-1299)discloses a method for the detection of adenovirus in eye fluidspecimens. The method comprises collecting a sample of eye fluid anddetecting the analyte on a paper disc by enzyme immunoadsorption. Thedetection, however, lacks specificity and sensitivity.

Thus, it is the objective of the invention to provide a sensitive andrapid non-invasive method for the detection of pathogens, e.g. bacterialor viral infectious agents in body fluids.

SUMMARY OF THE INVENTION

In a first aspect, the present invention relates to a method for thedetection of a target which is selected from pathogens and/orallergy-associated components in a body fluid comprising the steps:

-   (a) non-invasively collecting a body fluid sample with a swab    member,-   (b) transferring the sample to a application zone on a sample    analysis device and-   (c) analysing the sample.

In a further aspect, the invention relates to a method for diagnosingconjunctivitis comprising the steps:

-   (a) non-invasively collecting an eye fluid sample with a swab    member,-   (b) transferring the sample to a application zone on a sample    analysis device and-   (c) analysing the sample.

In still a further aspect, the invention relates to a test kitcomprising

-   (a) a swab member for non-invasively collecting a body fluid sample,-   (b) a sample analysis device comprising a detection zone, wherein    the detection zone contains reagents for determining the presence    and/or amount of at least one target which is selected from    pathogens and/or allergy-associated components.

In still a further aspect, the invention relates to a test kitcomprising

-   (a) a swab member for non-invasively collecting an eye fluid sample,-   (b) a sample analysis device comprising a detection zone, wherein    the detection zone contains reagents for determining the presence    and/or amount of at least one target which is selected from    pathogens and/or allergy-associated components wherein the target is    a causative agent or mediator of conjunctivitis or a plurality of    such causative agents and/or mediators.

DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a sample analysis device in the form of a chromatographictest strip comprising a plurality of different strip materials buildingan absorbent pad (1), an application zone (2), a detection zone (3) anda waste zone (4). The strip materials are arranged on an adhesiveplastic backing (5). The absorbent pad (1) is providing for adding anelution medium in order to facilitate the transfer of the sample to thedetection zone (3).

FIG. 2 shows a plastic housing (6) containing the strip as shown inFIG. 1. A sample application window (7) is provided for bringing a swabmember into contact with the strip. The test result is displayed in theread out window (8).

FIG. 3 shows a swab member or collection device for collecting a sample.The swab member comprises a plastic body (9) with a sample collectionmaterial (11) fixed on it and an opening (10) corresponding to a readout window when the swab member is operatively in contact with a teststrip.

FIG. 4 shows a test kit comprising a sample analysis device according toFIGS. 1 and 2 and a swab member according to FIG. 3.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

The invention provides a sensitive and rapid method for the detection oftargets, e.g. pathogens and/or allergy-associated components in samplescollected by non-invasive means from a body fluid. The pathogens areselected from viruses, microorganisms, e.g. bacteria and parasites, e.g.amoebae or nematodes. The allergy-associated components are selectedfrom allergens and anti-allergic components. The detection may comprisea direct detection of the target, e.g. the pathogen and/or the detectionof antibodies against the target, e.g. the pathogen which are present inthe fluid sample to be tested. Preferably, the method comprises aparallel determination of a plurality of targets.

The body fluid is preferably a fluid from a body surface selected frommucose membrane fluids (of the oral, nasal, vaginal, and ocularcavities) tears, secretions from glands and secretions from lesions orblisters, e.g. lesions or blisters on the skin. More preferably, thesample is selected from oral, nasal, ocular, genital and rectal fluidsand secretions from skin lesions or blisters. Most preferably, thesample is an eye fluid. A significant advantage of the method is thatresults are provided within the medical consultation period, e.g. in fewminutes. Preferably, the results are provided in a time period up to 20minutes, more preferably up to 15 minutes. Also, as the test isnoninvasive, it poses very little risk to the patient. Thus the bestavailable treatment can be applied on a timely basis for a specificpathogen. A further advantage over prior art methods is that only a fewmicroliters of sample are required to perform an analysis. The sample ispreferably about 0.1 μl to about 100 μl, more preferably about 0.2 μl toabout 20 μl and most preferably about 0.5 μl to about 10 μl.

The invention may be performed by means of a simple test kit. Handlingof the test kit does not necessitate additional laboratory equipment,further handling of reagents or instrumentation. Another importantadvantage of the invention described below is that the detection limitis typically 10 to 100 times lower than currently available diagnostictests because samples do not require dilution before they aretransferred to the analysis device. Therefore the disclosed method hasproven to be more sensitive and accurate than methods of the prior art.

The invention discloses a non-invasive method for the rapid andpoint-of-care determination of pathogens from body fluids. The method issuitable for diagnosis in human beings and animals, e.g. pets orlivestock animals. A preferred application is the detection of pathogensin eye fluid, e.g. human eye fluid. In this embodiment the pathogen tobe detected is a causative agent of conjunctivitis or a plurality ofsuch causative agents. For example, the pathogen is selected from thegroup of adenoviruses, herpesviruses, chlamydiae, cytomegaloviruses andcombinations thereof. More preferably, a plurality of pathogens aredetected on a single sample analysis device. For example, the sampleanalysis device may allow a simultaneous detection of a plurality ofpathogens, particularly of at least two, of at least three, of at leastfour or of at least five pathogens selected from the group consisting ofadenoviruses, herpesviruses, chlamydiae, cytomegaloviruses, pseudomonas,streptococci, haemophilus, staphylococci, amoebae, particularlyAcanthamoeba and nematodes, particularly Onchocera volvulus. Morepreferably, the method comprises a simultaneous detection ofadenoviruses, herpesviruses, chlamydiae, cytomegaloviruses andAcanthamoeba.

In addition the invention provides a non-invasive method for the rapidand point-of-care determination of at least one allergy-associatedcomponent, particularly an allergen (e.g. pollen, dust, etc.) and/or anantiallergen, particularly a component which is produced in the body inresponse to an allergenic challenge (e.g. IgE, histamine, etc.), in abody fluid as described above. More particularly, the invention relatesto methods and devices for the diagnosis of allergy-associatedcomponents in eye fluid, e.g. human eye fluid. In a preferredembodiment, the determination of at least one allergy-associatedcomponent may be combined with the determination of at least onepathogen as described above.

In the method of the invention, a body fluid sample is non-invasivelycollected with a collection device or swab member, respectively. Thecollection step preferably comprises wiping or dabbing the swab memberover a surface of the body containing body fluid to be tested. Usually,the swab member is sterile. The swab member may be dry or pretreatedwith a fluid before the collection step. For example, using a gentleswirling motion, a sterile swab member may be applied to the bodysurface or mucous membrane of concern and allowed to capture anypathogens and/or allergy-associated components contained in the bodyfluid.

The swab member may be a part which is separate from the sample analysisdevice and the sample is transferred by contacting the sample analysisdevice with the swab member under conditions wherein at least a part ofthe sample on the swab member is transferred to the sample analysisdevice. In this embodiment, the swab member is preferably contacted witha sample application zone on the analysis device from which the sampleis then transferred to the detection zone. The contact preferablycomprises fixing the swab member in a contact position with the sampleanalysis device in which the sample collection zone of the swab memberis in direct contact with the sample application zone of the analysisdevice. Thus, the swab member and/or the analysis device preferablycomprises fixing means for providing a fixed contact between both partsin a predetermined position. Alternatively, the swab member may be anintegrated part of the sample analysis device and the transfer comprisespassing at least a part of the sample on the swab member to thedetection zone on the sample analysis device.

The transfer of the sample from the swab member to the detection zone onthe sample analysis device is preferably a direct transfer, i.e. thetransfer takes place without pretreatment of the sample on the swabmember. Preferably, the transfer comprises an elution of the sample fromthe swab member with an elution medium, e.g. a buffer or water. Theelution medium may be added from an external source or may be providede.g. as a reservoir within the analysis device. Further, the transfer ispreferably a chromatographic and/or capillary transfer of fluid to thedetection zone on the sample analysis device.

In a preferred embodiment, the sample analysis device comprises achromatographic test strip, e.g. a lateral flow test strip. The sampleanalysis device may comprise a sample application zone, a detectionzone, optionally a waste zone, optionally a carrier backing, optionallya housing and optionally an opening for result read out. The sampleanalysis in the detection zone may be carried out by standard means,e.g. by an immunological or enzymatic detection method. Preferably, thedetection method comprises the use of test reagents capable ofspecifically binding the targets, e.g. pathogens to be tested orantibodies or other receptors against these targets, e.g. pathogens andsubsequent visualisation of the bound entity, e.g. by enzymaticdetection or by means of direct labelling groups, such as colloidalgold.

In an especially preferred embodiment, the swab member is placed on alateral flow test strip. With this step the collected specimen istransferred directly on an immunochromatographic or enzymatic teststrip. The test strip consists of one or several capillary activefleeces or membranes. The detection process will be either starteddirectly with sample transfer or may require an elution medium to beapplied for sample analysis. Preferably this elution medium is simpletap water. In the case of an immunochemical test strip, the chosenelution medium moves towards a detection zone and thereby passes thecontact site within the collection device. The analyte is diluted by theelution medium and carried with it to the detection zone. In thedetection zone the analyte is determined by qualitative and/orquantitative methods, e.g. in an immunological binding reaction.

The test strip can be made of one single chromatographic material, orpreferably several capillary active materials made of the same ordifferent materials and fixed on a carrier backing. These materials arein close contact with each other so as to form a transport path alongwhich a liquid driven by capillary forces flows from the start zone,passing the contact site of the swab and the detection zone, towards awaste zone at the other end of the strip.

Furthermore this invention is disclosing a device and test kit for theperformance of the described method.

In the method of invention, it is possible to make use of differentimmunological testing procedures to detect bacterial or viralconstituents on one or several immunological binding reactions. In apreferred embodiment, a chromatography test strip contains:

-   -   an application zone.    -   a conjungate zone containing at least one labeled binding        partner that is able to migrate with the elution medium. The        binding partner is capable of specifically binding to an analyte        and to a further specific reagent in the detection zone.    -   a detection zone containing a first section for the detection of        a first analyte, e.g. a test line, comprising an immobilized        specific binding partner for the analyte, and optionally further        sections for the detections of further analytes, and at least        one control section, e.g. a control line comprising an        immobilized specific binding partner of an indicator substance        indicating the functionality of the test kit.

In a preferred embodiment, the specific binding partners for theanalytes in the conjugate and the detection zone are monoclonal,polyclonal or recombinant antibodies or fragments of antibodies capableof binding to a pathogen. On the other hand, the specific bindingpartners may also be antigens capable of binding to antibodies against apathogen or an allergen. Other types of binding partners are bioorganicmacromolecules like aptamers or receptors. The conjugate zone may belocated before, within or after the sample application zone, seen in therunning direction of the eluent liquid. The test line(s) is(are) locatedafter the conjugate/application zone and the control line(s) is(are)located after the test line. Together, the test line(s) and controlline(s) comprise the detection zone.

Depending on the type of detection method, different binding partnersare present in the different zones. In a sandwich immunoassay, it ispreferred to have a labeled, non-immobilized analyte binding partner inthe conjugate zone. The binding partner forms a complex with the analytewhich is bound to the immobilized binding partner at the test line. In apreferred manner, the label of the conjugate binding partner is anoptically detectable label. Forming a complex at the test lineconcentrates and immobilizes the label and the test line gets visiblefor the bare eye, indicating a positive test result. Particularlypreferred are direct labels, and more particularly gold labels which canbe best recognized by the bare eye. Additionally, an electronicallyphotometrical read out device can be used to obtain more precise resultsand a semi-quantification of the analyte. Other labels may be latex,fluorophores or phosphorophores.

In order to test ocular fluids, a sample may be collected with a samplecollection device from the patient's eye by a health care professional.The sample collection device should be wiped or dabbed slightly severaltimes between in the inferior formix of the lower eye lid. If necessarythe collection device may be wet with sterile physiological saline todecrease patient's discomfort. This procedure is well known in theopthalmology practice as it is necessary for collecting specimens forconventional laboratory analysis. Generally the sample collection devicecomprises a capillary active material suitable for receiving a bodyfluid sample. In a preferred manner the sample collection material ismade out of fibers on the basis of cellulose, polyester, rayon orcalcium alginate. However, the sample collection device can also bedesigned as a microengineered mechanical structure containingmicrocapillaries and/or microchannels.

After the sample is collected, the collection device is fixed to theplastic housing containing the test strip (FIG. 4) and thereby thecollection applicator is slightly pressed on the application zone of thestrip. The collection device remains in this position.

In an alternative embodiment, the sample is taken by a standard swabmember as currently used in the physician's office or emergency rooms.This swab member is subsequently pressed into the application zone ofthe chromatographic test strip by means of an additional device similarto the sample collection unit.

In another preferred embodiment, the sample is taken by a swab memberand the sample collection devices is pressed for only a short time intothe application zone of the chromatographic test strip. A short periodof time preferably means a time up to 20 seconds, particularly between0.1 and 10 seconds. A transfer of the sample is happening within thecontact period.

In the next step, an elution medium is applied by dipping the absorbentpad into the chromatographic liquid. The absorbent pad is made of aparticularly well-absorbing material which delivers the liquid for theimmunochemical or enzymatic reactions. Preferred elution media are wateror buffer solutions that are conventionally used in immunoassays.

Alternatively the elution medium is contained in a reservoir which maybe integrated within the analysis device, e.g. as an ampoule or ablister. The reservoir may be opened by fixing the swab member or samplecollection device on the detection part of the device or by additionalmeans.

After a time period of up to 15 minutes, preferably within two to fiveminutes, the result can be read out in the detection zone. The result isconsidered positive when at least a partial area of the test line andthe control line shows a color change.

EXAMPLE

Test kit for the detection of adenovirus from patient's eye swab

The structure of a test strip is depicted in FIG. 1.

The polyester fleece for the absorbent pad was manufactured by Binzer,Hatzfeld, Federal Republic of Germany. The fleece is a polyester fleecereinforced with 10% curalon. The thickness ranges 1 and 2 mm, theabsorbance capacity is 1800 ml/m².

The application/conjugate zone consists of 80 parts polyester and 20parts viscous staple fibers at a thickness of 0.32 mm and an absorbingcapacity of 500 ml/m². The fleece is impregnated with the followingsolutions and then dried: 100 mmol/l HEPES Buffer, pH 7.5, 100 mol/lNaCl, conjugate of gold particles and anti-Hexon antibodies at aconcentration that has an optical density of 10 at 520 nm. Hexon is aprotein that is common in the capsid of human adenoviruses. The gold solwas manufactured according to standard procedures (Fres. Nature Vol.241, p. 20-22, 1973). Conjugation with the antibody was carried outaccording to prior art procedure (J. Immunol. Meth. Vol. 34, p. 11-31,1980). The sample application takes place in the application/conjugatezone.

The detection zone consists out of a nitrocellulose (NC) membrane with anominal pore size of 8 μm and a thickness of 100 μm produced bySchleicher & Schuell, Germany. The test line contains a Hexon specificantibody (not labeled) which is specific for a different epitope thanthe antibody immobilized on the gold. The control contains the sameantibody than the test line and binds any excess of Hexon specific gold.The control line will appear in any case even if Hexon is not presentindicating that the test worked correctly.

The chromatographic materials are in communication with each other inorder to create a fluid pathway.

A sample collection device is depicted in FIG. 3. The sample collectionmaterial may consist of bibulous material such as highly purified cottonfibers which are fixed to the plastic device by ultrasonic welding.Alternative materials may be polyester, rayon, polyamide or otherfibrous polymeric materials.

A test kit for the detection of Adenovirus antigen (as described in theexample above) was used in the Emergency Room of an OpthalmologicHospital to diagnose the clinical picture of a “pink” eye. From everypatient which has been tested with the test kit, a second sample wastaken and analysed in the laboratory.

The laboratory reference method used in this study was a combination ofcell culture and immunfluorescence (IF) detection (Rodrigues et al.,Opthalmology, 1979 March; 86(3):452-64) which is the current “laboratorygold standard” for determining the presence of adenovirus in humanocular fluid.

Within the testing period the following results have been achieved:

Cell Culture/IF + − Adeno test kit + 5 2 − 0 21

These preliminary results are equivalent to a diagnostic sensitivity of100% and a diagnostic specificity of 91%. These values are superior todiagnostic characteristics of other state of the art point-of-caredevices.

The invention claimed is:
 1. A method for the detection of a target which is selected from pathogens and/or allergy-associated components in a body fluid comprising the steps of: (a) non-invasively collecting a body fluid sample with a swab member; (b) directly contacting said swab member with a sample application zone on a lateral flow chromatographic test strip; wherein the sample is not diluted before contact between the swab member and the sample application zone on the lateral flow chromatographic test strip; and wherein at least a part of said body fluid sample is released directly into said sample application zone due to said swab member contacting said sample application zone; wherein the lateral flow chromatographic test strip further comprises a conjugate zone that is located at least partially within the sample application zone and comprises at least one binding partner for the target; (c) applying an elution medium to an absorbent pad on the lateral flow chromatographic test strip to transfer said sample to a detection zone on the lateral flow chromatographic test strip; and (d) analyzing the sample.
 2. The method of claim 1 wherein the pathogen is selected from viruses, bacterium, microorganisms and parasites.
 3. The method of claim 1 wherein the allergy-associated component is selected from allergens and anti-allergens.
 4. The method of claim 1 wherein the body fluid is fluid from a body surface selected from mucous membrane fluids, secretions from glands and secretions from lesions or blisters.
 5. The method of claim 4 wherein the sample is selected from oral, nasal, ocular, genital, and rectal fluids, and secretions from skin lesions or blisters.
 6. The method of claim 5 wherein the sample is an eye fluid.
 7. The method of claim 1 wherein the target is a pathogen which is a causative agent of conjunctivitis or a plurality of such causative agents.
 8. The method of claim 1 wherein the pathogen is selected from the group consisting of adenoviruses, herpesviruses, chlamydiae, cytomegaloviruses, pseudomonas, streptococci, haemophilus, staphylococci, amoebae and combinations thereof.
 9. The method of claim 1 wherein the target is an allergy-associated component which is a causative agent or a mediator of conjunctivitis or a plurality of such causative agents and/or mediators.
 10. The method of claim 1, wherein the target comprises at least one pathogen and at least one allergy-associated component.
 11. The method of claim 1 wherein the sample is about 0.1 μl to about 100 μl.
 12. The method of claim 11 wherein the sample is about 0.5 μl to about 10 μl.
 13. The method of claim 1 wherein step (a) comprises the substep of wiping or dabbing the swab member over a surface of a body part containing the body fluid to be tested.
 14. The method of claim 1 wherein the swab member is sterile.
 15. The method of claim 1 wherein the swab member is a part separate from the lateral flow chromatographic test strip.
 16. The method of claim 1 further comprising the step of fixing the swab member in a contact position with the lateral flow chromatographic test strip.
 17. The method of claim 1 wherein the elution medium is selected from the group consisting of a buffer or water.
 18. The method of claim 1 wherein the elution medium is added from an external source or is provided within the lateral flow chromatographic test strip.
 19. A method of claim 1 wherein step (d) comprises an immunological or enzymatic detection.
 20. The method of claim 1, wherein the binding partner in the conjugate zone comprises at least one labeled binding partner that is able to migrate with the elution medium.
 21. A method for the detection of a target which is selected from pathogens and/or allergy-associated components in a body fluid comprising the steps of: a) non-invasively collecting a body fluid sample with a swab member; b) directly contacting said swab member and said sample on said swab member with a sample application zone on a lateral flow chromatographic test strip, wherein dilution of the sample does not occur before direct contact between the swab member and the sample and the sample application zone on the lateral flow chromatographic test strip; c) applying an elution medium to an absorbent pad on said lateral flow chromatographic test strip to transfer said sample to a detection zone on the chromatographic test strip; and d) analyzing the sample; wherein the lateral flow chromatographic test strip further comprises a conjugate zone located at least partially within the sample application zone, wherein said conjugate zone comprises at least one labeled binding partner that is able to migrate with the elution medium wherein, when the analyte is present, the labeled binding partner complexes with the analyte and then migrates to the detection zone.
 22. The method of claim 21, wherein at least a part of said body fluid sample is released directly into said sample application zone due to said swab member contacting said sample application zone.
 23. The method of claim 21, wherein the body fluid sample is an eye fluid sample.
 24. The method of claim 21, wherein the target is a pathogen which is a causative agent of conjunctivitis or a plurality of such causative agents.
 25. A method for the detection of a target which is selected from pathogens and/or allergy-associated components in a body fluid comprising the steps of: a) non-invasively collecting a body fluid sample with a swab member; b) directly contacting said swab member with a sample application zone on a lateral flow chromatographic test strip, wherein at least a part of said body fluid sample is released directly into said sample application zone due to said swab member contacting said sample application zone; and wherein the lateral flow chromatographic test strip further comprises a conjugate zone that is located at least partially within the sample application zone and comprises at least one labeled binding partner for the target; and c) applying an elution medium to an absorbent pad on the lateral flow chromatographic test strip to transfer said sample to a detection zone on the lateral flow chromatographic test strip; wherein the sample is not diluted prior to steps b) and c); and d) analyzing the sample.
 26. A method for the detection of a target which is selected from pathogens and/or allergy-associated components in a body fluid comprising the steps of: a) non-invasively collecting a body fluid sample with a swab member; b) directly contacting said swab member and said sample on said swab member with a sample application zone on a lateral flow chromatographic test strip; and wherein the lateral flow chromatographic test strip further comprises a conjugate zone that is located at least partially within the sample application zone and comprises at least one labeled binding partner for the target; c) applying an elution medium to an absorbent pad on the lateral flow chromatographic test strip to transfer said sample to a detection zone; wherein the sample is not diluted prior to steps b) and c); and d) analyzing the sample.
 27. A method for the detection of a target which is selected from pathogens and/or allergy-associated components in a body fluid comprising the steps of: a) non-invasively collecting a body fluid sample with a swab member; b) directly contacting said swab member and said sample on said swab member with a sample application zone on a lateral flow chromatographic test strip, wherein dilution of the sample does not occur before direct contact between the swab member and the sample and the sample application zone on the lateral flow chromatographic test strip; c) applying an elution medium to an absorbent pad on said lateral flow chromatographic test strip to transfer said sample to a detection zone on the chromatographic test strip; and d) analyzing the sample; wherein the lateral flow chromatographic test strip further comprises a conjugate zone located within the sample application zone, wherein the conjugate zone comprises at least one labeled binding partner that is able to migrate with the elution medium, and wherein, when the analyte is present, the labeled binding partner complexes with the analyte and then migrates to the detection zone. 